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Three‐Dimensional Behavior of Ice Crystals and Biological Cells during Freezing of Cell Suspensions a
Author(s) -
ISHIGURO HIROSHI,
KOIKE KAZUHITO
Publication year - 1998
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.1998.tb10157.x
Subject(s) - acridine orange , ice crystals , suspension (topology) , confocal , biophysics , liquid crystal , materials science , suspension culture , ice formation , chemistry , cell culture , optics , biology , biochemistry , apoptosis , physics , mathematics , optoelectronics , genetics , homotopy , pure mathematics , atmospheric sciences , geology
Behavior of ice crystals and human red blood cells during extracellular‐freezing was investigated in three‐dimensions using a confocal laser scanning microscope(CLSM), which noninvasively produces tomograms of biological materials. Physiological saline and physiological saline with 2.4M glycerol were used for suspension. Various cooling rates for directional solidification were used for distinctive morphology of the ice crystals. Addition of acridine orange as a fluorescent dye into the cell suspension enabled ice crystal, cells and unfrozen solution to be distinguished by different colors. The results indicate that the microscopic structure is three‐dimensional for flat, cellular, and dendritic solid‐liquid interfaces and that a CLSM is very effective in studying three‐dimensional structure during the freezing of cell suspensions.